Method and application for testing and verifying SNPs (single nucleotide polymorphisms) polymorphic site hereditary effects in CIDEC gene 5 end control region

Abstract

The invention discloses a method for testing and verifying cattle CIDEC gene SNPs (Single Nucleotide Polymorphisms) site hereditary effects, which comprises the following steps of: by taking cattle whole genomic DNA containing CIDEC gene as a template and a primer pair P as a primer, carrying out PCR (Polymerase Chain Reaction) amplification on the different cattle CIDEC gene haplotypes; digesting PCR amplification products and pGL3-Basis empty gsensor by using restriction enzyme KpnI and XhoI respectively, then connecting the different haplotypes to pGL3-Basis carrier by using DNA ligase to establish pGL3-Basic-CIDEC-H1,H2,H3,H4,H8 and H9 expression vectors; transfecting the established expression vectors in 3T3-L1 cells; using a bifluorescence reporting system kit to detect the transcriptional activity of different haplotypes; and testing and verifying the association analysis result by combining the transcriptional activity of different haplotypes and the information that the SNP site changes a cis acting element. The detection method disclosed by the invention lays a basis for test and verification of CIDEC gene single nucleotide polymorphism hereditary effects, so as to be used for marker assisted selection (MAS) of cattle meat growth trait in China, and quickly establish cattle population with excellent genetic resources.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and relates to a verification method and application of the genetic effect of a single nucleotide polymorphism (SNP) site in a gene, in particular to a method for verifying the genetic effect of a single nucleotide polymorphism site in a cattle CIDEC gene Authentication method. Background technique [0002] SNP is a genetic marker formed by a single nucleotide variation on the genome. There are many SNPs on the genome and polymorphisms are abundant. According to research, SNPs are quite widely distributed in the genome, and appear once every 300 base pairs in the human genome. SNPs can be classified as single nucleotide substitutions, transversions, deletions and insertions. From the perspective of the molecular mechanism of the influence on the genetic traits of organisms, SNP can be divided into two types: one is synonymous cSNP (synonymousc SNP), that is, the change of the coding sequence c...

AI Technical Summary

Problems solved by technology

At present, there are many studies on SNPs in coding regions. For SNPs in non-coding regions, although their functional research is of great significance, SNP data in this region are difficult to collect, so they have not received the same attention as the research on SNPs in coding regions.

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